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Nucleic Acids Research Advance Access published online on December 17, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm992
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Red flag on the white reporter: a versatile insulator abuts the white gene in Drosophila and is omnipresent in mini-white constructs

Darya Chetverina1, Ekaterina Savitskaya1,2, Oksana Maksimenko1, Larisa Melnikova1, Olga Zaytseva1, Alexander Parshikov1, Alexander V. Galkin3 and Pavel Georgiev1,*

1Department of the Control of Genetic Processes, Institute of Gene Biology, Russian Academy of Sciences, 2University of Oslo, Centre for Medical Studies in Russia, Moscow 199334, Russia and 3Department of Molecular Immunogenetics of Cancer, Institute of Gene Biology, Russian Academy of Sciences, Moscow 119334, Russia

*To whom correspondence should be addressed. Tel: 7 499 1359734; Fax: 7 499 1354105; Email: georgiev_p{at}mail.ru

Received September 4, 2007. Revised October 22, 2007. Accepted October 22, 2007.

Much of the research on insulators in Drosophila has been done with transgenic constructs using the white gene (mini-white) as reporter. Hereby we report that the sequence between the white and CG32795 genes in Drosophila melanogaster contains an insulator of a novel kind. Its functional core is within a 368 bp segment almost contiguous to the white 3'UTR, hence we name it as Wari (white-abutting resident insulator). Though Wari contains no binding sites for known insulator proteins and does not require Su(Hw) or Mod(mdg4) for its activity, it can equally well interact with another copy of Wari and with unrelated Su(Hw)-dependent insulators, gypsy or 1A2. In its natural downstream position, Wari reinforces enhancer blocking by any of the three insulators placed between the enhancer and the promoter; again, Wari–Wari, Wari–gypsy or 1A2–Wari pairing results in mutual neutralization (insulator bypass) when they precede the promoter. The distressing issue is that this element hides in all mini-white constructs employed worldwide to study various insulators and other regulatory elements as well as long-range genomic interactions, and its versatile effects could have seriously influenced the results and conclusions of many works.

Present address: Ekaterina Savitskaya, Centre of Bioengineering, Russian Academy of Sciences, Prosp. 60-letiya Oktyabrya, bldg. 7-1, Moscow 117312, Russia


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